Molecular cloning and expression of anti-tumor adhesion peptide (beta3).
- Author:
Song-Mei WANG
1
;
Jun ZHU
;
Yan LI
;
Luan-Feng PAN
;
Xi-Liang ZHA
;
Yin-Kun LIU
Author Information
1. Laboratory of Molecular Biology, Shanghai Medical College, FuDan University, Shanghai 200032, China.
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Antineoplastic Agents;
pharmacology;
Cell Adhesion;
drug effects;
Cloning, Molecular;
Escherichia coli;
genetics;
metabolism;
Molecular Sequence Data;
Peptide Fragments;
pharmacology;
Peptides;
genetics;
metabolism;
pharmacology;
Recombinant Proteins;
biosynthesis;
genetics;
Tumor Cells, Cultured
- From:
Chinese Journal of Biotechnology
2005;21(4):558-562
- CountryChina
- Language:Chinese
-
Abstract:
To block tumor cell adhesion, inhibit tumor metastasis and recurrence, the anti-adhesion peptide-trimeric beta peptide (DLYYLMDLSYSMKGGDLYYLMDLSYSMKGGDLYYLMDLSYSMK, beta3) was designed. The DNA fragment of beta3 was cloned into expression vector pET-His and the fusion protein His-beta3 was expressed in E. coli. BL21(DE3)plysS. After 1.5 hours' induction with IPTG, His-beta3 peptide was expressed significantly amounting to 10% of the insoluble proteins and 4% of the total proteins. 20mg of beta3 peptide was obtained from one litter culture medium after purification by using metal-chelating sepharose 6B FF. The purity of beta3 is 92.2% according to Gel-Pro analysis. The anti-adhesion effects of beta3 peptide, beta1 peptide (DLYYLMDLSYSMK) and GRGDS on the hepatocellular carcinoma cell line SMMC-7721 and the high metastasis hepatocellular carcinoma cell line HCCLM6 were studied. The result showed the beta3 blocked the adhesion of HCCLM6 cells and SMMC-7721 cells to fibronectin (FN) specifically. The inhibition effect was dose-dependent and time-dependent and the inhibition rate of beta3 was higher than three times concentration of beta1 and GRGDS. This suggested that pET-His-beta3/BL21(DE3)plysS was a suitable expression system for beta3, and the expressed beta3 specially inhibited the adhesion of cancer cells.
